目的 探讨长链非编码RNA LINC00963通过调控TLR4/MyD88/NF-κB信号通路减轻脓毒症心肌损伤(SIMD)的作用机制。方法 采用盲肠结扎穿孔术(CLP)建立脓毒症小鼠模型(n=24),随机分为LINC00963过表达慢病毒组(LV-LINC00963-OE组)、敲低组(LV-LINC00963-KD组)及相应阴性对照组。通过慢病毒载体转染RAW264.7巨噬细胞,经尾静脉注射调控LINC00963表达,术后7 d取心脏组织行HE染色评估病理损伤程度;采用Western blot检测TNF-α、NF-κB p65、MyD88及AP-1蛋白表达。结果 LINC00963敲低组心肌间质水肿评分显著高于对照组[(3.8±0.4) vs. (1.2±0.3)],P<0.01,且促炎因子TNF-α、NF-κB p65、MyD88及AP-1表达上调1.7~2.5倍,差异有统计学意义(P＜0.01);在LINC00963过表达组中,上述指标显著降低,差异有统计学意义(P＜0.01)。结论 LINC00963通过靶向抑制TLR4/MyD88/NF-κB信号通路介导的炎症反应,减轻脓毒症心肌损伤,为临床干预脓毒症心肌病提供了潜在治疗靶点。
Abstract
Objective To explore the mechanism of long non-coding RNA LINC00963 in alleviating myocardial injury in sepsis through regulating the TLR4/MyD88/NF-κB signaling pathway. Methods A murine sepsis model was established by cecal ligation and puncture (CLP) (n=24). Mice were randomly divided into a LINC00963 overexpression lentivirus group (LV-LINC00963-OE), a LINC00963 knockdown lentivirus group (LV-LINC00963-KD), and a corresponding negative control group. RAW264.7 macrophages were transfected with lentiviral vectors, and the LINC00963 expression was regulated by tail vein injection. Cardiac tissues were harvested 7 days after surgery for HE staining to evaluate the degree of pathological injury; Western blot was used to detect the expression of TNF-α, NF-κB p65, MyD88, and AP-1 proteins. Results The myocardial interstitial edema score in the LV-LINC00963-KD group was significantly higher than that in the control group [(3.8±0.4) vs. (1.2±0.3), P<0.01], and the expression of pro-inflammatory factors TNF-α, NF-κB p65, MyD88, and AP-1 was upregulated by 1.8 to 2.5 times, with statistically significant differences (P<0.01). Conversely, in the LV-LINC00963-OE group, the above indicators were significantly reduced , with statistically significant differences (P<0.01). Conclusions LINC00963 mediates the inflammatory response by targeting the TLR4/MyD88/NF-κB signaling pathway, alleviating myocardial injury in sepsis, providing a potential therapeutic target for clinical intervention of septic cardiomyopathy.
关键词
脓毒症心肌病 /
LINC00963 /
促炎细胞因子 /
TLR4/MyD88/NF-κB通路 /
细胞凋亡
Key words
septic cardiomyopathy /
LINC00963 /
pro-inflammatory cytokines /
TLR4/MyD88/NF-κB signaling pathway /
apoptosis
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